Recognition of the A chain carboxy-terminal heparin binding region of fibronectin involves multiple sites: two contiguous sequences act independently to promote neural cell adhesion.
Cellular interactions with fibronectin-treated substrata have a complex molecular basis involving multiple domains. A carboxy-terminal cell and heparin binding region of fibronectin (FN) is particularly interesting because it is a strong promoter of neurite outgrowth (Rogers, S.L., J.B. McCarthy, S.L. Palm, L.T. Furcht, and P.C. Letourneau, 1985. J. Neurosci. 5:369-378) and cell attachment (McCarthy, J.B., S.T. Hagen, and L.T. Furcht. 1986. J. Cell Biol. 102:179-188). To further understand the molecular mechanisms of neuronal interactions with this region of FN, we screened two peptides from the 33-kD heparin binding fragment of the FN A chain, FN-C/H II (KNNQKSEPLIGRKKT) and CS1 (Humphries, M.J., A. Komoriya, S.K. Akiyama, K. Olden, and K.M. Yamada. 1987. J. Biol. Chem. 262:6886-6892), for their ability to promote B104 neuroblastoma cell-substratum adhesion and neurite outgrowth. Both FN-C/H II and CS1 promoted B104 cell attachment in a concentration-dependent and saturable manner, with attachment to FN-C/H II exceeding attachment to CS1. In solution, both exogenous FN-C/H II or CS1 partially inhibited cell adhesion to the 33-kD fragment. Similar results were obtained with anti-FN-C/H II antibodies. In contrast, soluble GRGDSP did not affect B104 cell adhesion to FN-C/H II. These results indicate that both FN-C/H II and CS1 represent distinct, RGD-independent, cell adhesion-promoting sites active within the 33-kD fragment, and further define FN-C/H II as a novel neural recognition sequence in FN. B104 adhesion to FN-C/H II and CS1 differs in sensitivity to heparin, yet each peptide inhibited adhesion to the other peptide, suggesting cell adhesion is somehow related at the cellular level. Within the A chain 33-kD fragment, FN-C/H II and CS1 are contiguous, and might represent components of a larger domain with greater neurite-promoting activity since only the 33-kD fragment, and neither individual peptide, was effective at promoting B104 neurite outgrowth. These data further support the hypothesis that cell responses to FN are mediated by multiple sites involving both heparin-sensitive and -insensitive mechanisms.
Haugen
P K
PK
Department of Cell Biology, University of Minnesota, Minneapolis 55455.
McCarthy
J B
JB
Skubitz
A P
AP
Furcht
L T
LT
Letourneau
P C
PC
D000118:Acetylglucosaminidase / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D001665:Binding Sites; D002448:Cell Adhesion; D002460:Cell Line; D005353:Fibronectins / Q000378:metabolism* / Q000502:physiology; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D008969:Molecular Sequence Data
D000906:Antibodies; D005353:Fibronectins; D046911:Macromolecular Substances; D006493:Heparin; D000118:Acetylglucosaminidase